环状RNA circ_0001821通过exosomes播散NSCLC化疗耐药性的机制研究

Circular RNAs (circRNAs) represent a new class of non-coding RNAs that is involved in the development of cancer. Here, we identified circ_0001821 as one circRNA significantly upregulated in NSCLC resistant cells and associated with chemotherapeutic sensitivity of NSCLC cells. Furthermore, circ_0001821 could be incorporated into exosomes and transmitted to sensitive cells. Target gene prediction indicates that mir-192, mir-193b and mir-152 may be regulated by circ_0001821. Based on the above research, this project is proposed to carry out four aspects of research: (1) Using functional gain and loss experiments, determine circ_0001821 biology functions in NSCLC chemoresistance; (2) Using bioinformatics, qPCR and RIP, screen circ_0001821 function related key regulatory molecules; (3) Analysis of the effect of exosome-transmitted circ_0001821 on disseminating resistance in vitro and in vivo; (4) Analyze the potential diagnostic values of circulating exosomal circ_0001821 in NSCLC. This study will provide new valuable resistance molecular markers and therapeutic targets for NSCLC.

环状RNA（circRNAs）在肿瘤发生发展中发挥重要作用。课题组前期发现异常表达的环状RNA circ_0001821与非小细胞肺癌化疗耐药相关，并能通过外泌体（exosomes）传递进入敏感细胞。靶基因预测表明miR-192、miR-193b和miR-152等多个具有抑癌作用的miRNAs可能受到circ_0001821的调控。为此，本课题拟（1）功能获得和缺失验证circ_0001821在NSCLC 耐药中的作用；（2）生物信息学、qPCR和RIP等方法确定circ_0001821调控耐药的靶基因；（3）体内外分析exosomes传递circ_0001821播散NSCLC耐药性的作用;（4）检测循环exosomal circ_0001821的表达，分析其对NSCLC诊断和化疗反应预测的价值。该工作有望为临床个体化治疗NSCLC提供新的耐药分子标记物和治疗靶点。

本研究通过高通量环状RNA（circRNAs）芯片对非小细胞肺癌（NSCLC）多西他赛耐药细胞系A549/DTX，H1299/DTX，及临床上以多西他赛为基础化疗的敏感和复发组织标本进行差异表达谱进行分析，发现环状RNA circ_0001821在耐药细胞和组织中表达上调，体内外功能实验证实circ_0001821能够调控NSCLC细胞对多西他赛的敏感性，高表达的circ_0001821能够通过外泌体传递进入受体细胞，进而播撒耐药性。机制上，circ_0001821 充当 miR-148a-3p 的海绵，而miR-148a-3p能够与SLC2A1 3′UTR 作用。Circ_0001821 通过 miR-148a-3p 靶向调节 SLC2A1 表达促进NSCLC细胞多西他赛耐药。此外，circ_0001821在肺癌血浆中的表达显著升高，且与肺癌患者临床分期相关。ROC曲线评估显示，circ_0001821对肺癌诊断的曲线下面积为0.792 (95% CI 0.724-0.850)，联合肺癌的传统标志物NSE，能显著提高对肺癌的诊断效率。这些结果提示circ_0001821是肺癌潜在的治疗靶点和诊断标志物。